Molecular and General Genetics MGG

, Volume 200, Issue 2, pp 328–334 | Cite as

Molecular cloning and functional analysis of the cysG and nirB genes of Escherichia coli K12, two closely-linked genes required for NADH-dependent nitrite reductase activity

  • Heather Macdonald
  • Jeff Cole
Article

Summary

We have cloned two genes, nirB+and cysG+which are required for NADH-dependent nitrite reductase to be active, from the 74 min region of the Escherichia coli chromosome. Restriction mapping and complementation analysis establish the gene order crp-nirB-cysG-aroB. Both genes are trans-dominant in merodiploids and, under some conditions, can be expressed independently. The cysG+gene can be expressed from both high and low copy number plasmids carrying a 3.6 kb PstI-EcoRI restriction fragment. Attempts to sub-clone the nirB+gene into pBR322 on a 14.5 kb EcoRI fragment were unsuccessful, but this fragment was readily sub-cloned into and expressed from the low copy number plasmid pLG338 (Stoker et al. 1982). Overproduction of the 88 kDa nitrite reductase apoprotein by strains carrying a functional nirB+gene suggests that nirB is the structural gene for this enzyme.

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Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • Heather Macdonald
    • 1
  • Jeff Cole
    • 1
  1. 1.Department of BiochemistryUniversity of BirminghamBirminghamU.K.

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